BACKGROUND:The successful establishment of human embryonic stem cel lines in vitro is of great significance to human embryonic development mechanism and developmental biology, cel and tissue transplantation in the treatment of certain diseases. OBJECTIVE:To summarize the progress of in vitro culture of embryos and establishment of embryonic stem cel lines, to explore the influential factors for in vitro culture of embryos, and the methods of culturing human discarded embryos, isolating inner cel mass and establishing embryonic stem cel lines, as wel as the establishing conditions for embryonic stem cel lines. METHODS:With the key words of“embryo, embryonic stem cel s, coculture, sequential culture”, the first author searched CNKI and SCI databases for literatures concerning in vitro culture and transplantation of embryos and establishment of embryonic stem cel lines published from 2000 to 2014. Systematic evaluation was conducted. Final y, 58 literatures were retained for result analysis. RESULTS AND CONCLUSION:The culturing condition for embryos in vitro is the key factor affecting embryo transfer outcomes, including culture medium component and culture system. In previous studies, the component and application of culture medium have changed greatly, and the culture system has altered from single culture to coculture and sequential culture. Ethical issues and embryonic origin restrictions restrict the establishment of human embryonic stem cel lines. Clinical y discarded low-quality embryos can be used as one of the material sources to establish human embryonic stem cel lines, which can effectively lessen the problem of embryo shortage during the establishment of human embryonic stem cel lines and reduce ethical disputes.

BACKGROUND: Early mammalian embryonic development and implantation is a very complex process, which is regulated by a variety of cytokines. Moreover, epidermal growth factor, transforming growth factor, leukemia inhibitory factor, insulin-like growth factor and vascular endothelial growth factor play an important role in this regulation process.
OBJECTIVE: To introduce cytokines related to early embryonic development, including types, structural features and molecular mechanisms, and meanwhile to review the effects of epidermal growth factor, transforming growth factor, leukemia inhibitory factor, insulin-like growth factor and vascular endothelial growth factor in the early embryonic development.
METHODS:A literature retrieval was performed to search relevant articles published from 1989 to 2015 using the keywords of “embryos development, epidermal growth factor, transforming growth factor, leukemia inhibitory factor, insulin-like growth factor, vascular endothelial growth factor”. Active effects of these five kinds of cytokines in overcoming the mammalian embryonic development block were reviewed and summarized to provide theoretical and experimental basis for further optimization of the embryonic culture system in vitro.
RESULTS AND CONCLUSION: Cel growth factors involved in embryonic development is the focus of the study. Insulin-like growth factor is capable of promoting the embryonic growth; epidermal growth factor exhibits multiform functions in the embryonic development; leukemia inhibitory factor directly affects oocyte cel development, and progesterone increases the expression of leukemia inhibitory factor to provide a good cytokine microenvironment for embryonic development; vascular endothelial growth factor promotesin vitro oocyte maturation, and plays a very important role in various stages of embryonic development. To clarify these regulatory patterns of cytokines cannot only contribute toin vitro embryonic culture, but also reduce embryonic apoptosis, which is of great significance for promoting embryonic development.

Objective To report the clinical results of the treatment for recurrent immunologic disorders of cornea and conjunctiva by conjunctival resection with amniotic membrane transplantation(AMT). Design A retrospective clinical case series. Participants Five patients 5 eyes with recurrent immunologic disorders of cornea and conjunctiva were enrolled into this study, including 2 eyes of phlytenular keratoconjunctivitis with corneal lesion, 1 eye of nodular episcleritis, 2 eyes of corneoscleral interstitial keratitis. Methods Patients were treated with partial conjunctiva resection and AMT. After operation, steroids and cyclosporin eye-drops were given for 2-3 weeks, and patients were followed up for 6-12 months. Main Outcome Measures Complications and recurrent situations were observed. Results The corneal disease kept quiescent, and the blood vessel subsidized. There were no recurrences during the follow up period in 4 cases. Recurrent signs were noted in one patient at 12 months after surgery. But no recurrence of corneal lesion was noted although the conjunctivitis was severe. Conclusion Local conjunctiva resection combined with AMT is a useful method for recurrent cases of immunologic disorders of cornea and conjunctiva. (Ophthalmol CHN, 2006,15:177-179)

Background and purpose: Docetaxel/cisplatin are widely used in chemo-naive patients with advanced non-small cell lung cancer(NSCLC),but the standard 3-weeks'project of docetaxel caused significant toxicity.We performed this study to compare the effect and toxicity of modified and standard 3-weeks'docetaxel/cisplatin as first line chemotherapy for advanced NSCLC.Methods:68 patients with stage ⅢB or Ⅳ NSCLC(proven by histology or cytology) were randomly divided into two groups,modified(A) and standard(B) chemotherapy.Group A: docetaxel 75 mg/m2,divided into 2 days,ivgtt d1 and d 8,cisplatin 25 mg/(m2?d),ivgtt d 1-d 3,q3w;Group B: docetaxel 75 mg/m2,ivgtt d 1,cisplatin was administered as Group A,q3w.The effect and toxicity were assessed after two cycles and one-year survival was followed up.Results:There was no CR in both groups.10 PR,20 SD,4 PD were found in group A,the overall response rate is 29%;whereas 11 PR,20 SD,3 PD were found in group B,the overall response rate is 32%.The one-year survival rate were 38% and 35% in group A and B,respectively.There were no significant difference about the overall response rate(P=0.793) and one-year survival rate(P=0.801) between group A and B.The rates of grade Ⅲ/Ⅳ neutropenia were 18% and 47% in group A and B respectively.The difference was statistically significant(P=0.010).Conclusions:In comparison with the standard 3-weeks'docetaxel/ cisplatin chemotherapy,the modified one has similar response rate but lower hematologic toxicity,and thus it was well tolerated.

Objective To investigate effects of neural retina on development of the structure of outer blood retinal barrier in embryogenesis. Methods The retinal neural epithelium (RNE) and pigment epithelium (RPE) layers of 150, 120 and 90 embryonic chicken eyes incubated for 7, 10, and 14 days were peeled off. RNE was used to prepare the culture medium with different conditions (7drcSF3, 10drcSF3, 14drcSF3). RPE cells of 7- and 14-incubated chicken embryos were cultured on laminin-coated transwell filter. The SF3, 7drcSF3, 10drcSF3, 14drcSF3 medium were used respectively in the apical chamber and SF2 was used in basolateral chamber. After the formation of monolayer, the transepithelial electrical resistance of the RPE was detected. After the fixation of RPE cells, the condition of the tight junction among the cells was observed by immunohistochemistry and transmission electron microscopy. Results For the RPE cells of 7- and 14-day incubated embryonic eyes, the difference of TER in various medium of SF3/SF2, 7drcSF3/SF2, 10drcSF3/SF2, 14drcSF3/SF2 was statistically significant (P

Objective To compare the results of three different types of fine particulate matter laser light scattering aerosol monitors and to explore the influencing factors.Methods Three types of Aerosol Monitor A, B, and C were used to monitor the mass concentration of indoor PM2.5 at the same time and place and the membrane weighing method was used at the same time.In order to explore the possible impacts of temperature and humidity indoor on the results of monitoring, a simultaneous monitoring on the temperature and humidity indoor was conducted.The sampling continued 2 days and with 23 hours per day.Results The results of PM2.5 monitored by the three types of aerosol monitors had a good consistency(r were 0.965, 0.988, and 0.984 respectively), but there was significant difference among them(P

Objective To explore the effect of a herbalcompound Gehua Jiejue Dizhi Decoction (GJDD) on the liver fat deposition and the expression of PXR, and the mRNA and protein expression of its target genes CYP3A11 and CYP3A25in the liver tissues of mouse models of alcoholic fatty liver.Methods Twenty-nine healthy male C57BL/6J mice were randomly divided into control group (n=5), model group (n=8), high dose GJDD group (n=8)and low dose GJDD group (n=8).The mouse model of alcoholic fatty liver was prepared according to the National Institute on Alcohol Abuse and Alcoholism (NIAAA) method.Then, the mice were treated with the high dose and low dose GJDD for 9 days.Serum glutamic-pyruvic transaminase (AST) and aspartate aminotransferase (AST) were detected by enzyme-linked immunosorbent assay (ELISA).Liver fat deposition was detected by oil red O staining.Real-time RT-PCR and immunohistochemistry were performed to examine the expressions of PXR, CYP3A11 and CYP3A25.Results Compared with the model group, the liver fat deposition in the intervention groups was significantly reduced in a dose-dependent manner, with a significant increase of the expression of PXR and CYP3A25 (P < 0.01).The serum ALT level was significantly reduced in the model group (P < 0.01), while the transcriptional levels of CYP3A11 mRNA in the groups were similar (P ≥ 0.05).Conclusions Gehua Jiejue Dizhi Decoction has obvious therapeutic effect on the AFLD in mice, which may be related to the activation of PXR and its target genes CYP3A25.

Objective To research the relationship between the expression of PAI-1 and the clinical characteristics of the endometrial carcinoma.Methods We detected the level of the serum PAI-1 by ELISA in the patients with endometrial carcinoma,the patients with endometrial hyperplasia and the patients with normal endometrium.The expression of PAI-1 in endometrial carcinoma and normal endometrial tissues was observed by immunohistochemistry.Results The concentration of serum PAI-1 in patients with endometrial carcinoma was 18.64 ± 6.22 μtg/L,significantly higher than those of patients with endometrial hyperplasia (6.94 ± 2.87) μg/L and patients with normal endometrium (6.68 ± 2.13)μg/L (P=0.00).The expression rate of PAI-1 was 68.2％ (15/22) in endometrial carcinoma tissue,and 8.3％ (2/24) in normal endometrial tissue (P=0.00).Compared with the early endometrial carcinoma,the expression rate of PAI-1 in the advanced endometrial carcinoma was significantly higher (6/6 w 9/16,P=0.03).Conclusions The expression level of PAI-1 may be related to the stage of the endometrial carcinoma.

Objective The aim of this study was to observe the growth difference and expression of cytochrome C of skin hair follicles in neonatal mice .Methods The morphology of different skin hair follicles of neonatal mice ( postnatal day 1-9)were observed by HE staining histology and cytochrome C was detected by immunohistochemistry .Results The skin hair follicles in different parts of neonatal mice showed differences not only in morphology but also in developmental pe -riods.Hair follicle growth in the back and tail skin had a nonlinear and growing period .After the nonlinear and growing pe-riod they began to grow rapidly .The tail development was slightly slower than that on the back .The hair follicles of vibris-sae were very special , and started to develop without a stable period .Conclusions The results of morphological observa-tion and cytochrome C immunohistochemistry demonstrate that differences exist in the hair follicle morphology and develop -mental times in the skin of different parts of the body in neonatal mice .

Objective To investigate the dynamic changes of endoplasmic reticulum stress-related molecules including glucose regulated protein (GRP78), C/EBP homologous protein (CHOP), and caspase-12 in sciatic nerve of diabetic rats and explore its mechanisms.Methods Rats were randomly divided into normal control group (NC) and diabetes mellitus group (DM) that were induced by intraperitoneal injection of Streptozocin after 4 weeks of high-fat chow feeding.Sciatic nerves were isolated for three times at 4 weeks, 8 weeks and 12 weeks after induction of diabetes.The expressions of GRP78, CHOP,and caspase-12 were detected with quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analyses.The morphology of sciatic nerve was investigated with electron microscope.Results With the extension of the course, demyelinating and axonal injury appeared in sciatic nerve of diabetic rats.The expressions of GRP78 mRNA and protein in DM group were significantly higher than NC group at 4 weeks and 8 weeks after induction of diabetes(P ＜0.05, P ＜0.01).The expressions of CHOP mRNA and protein in DM group were significantly higher than NC group at 8 weeks and 12 weeks after induction of diabetes (P ＜ 0.05).The expressions of caspase-12 mRNA and protein in DM group were significantly higher than NC group at 8 weeks after induction of diabetes(P ＜ 0.05, P ＜ 0.01).Conclusions Endoplasmic reticulum stress-related molecules (GRP78, CHOP, and caspase-12) contributed to the peripheral nerve injury of diabetic rats, and displayed dynamic changes.